Method for producing a plant extract and fraction, pharmaceutical compositions and use thereof

Abstract

The present invention describes the obtaining process of the dichloromethane fraction and subfractions from Eleutherine plicata , popularly known as “marupazinho, marupari, palmeirinha, coquinho, marupaí, marupaú, marupá-piranga, and lirio-folha-de-palmeira”. This invention comprises the obtaining process of pharmaceutical compositions that contain the dichloromethane fraction and/or naphthoquinone, as well as its use for malaria treatment. Extract and fractions were assayed and presented antiplasmodial activity, particularly against chloroquine-resistant Plasmodium falciparum (clone W2).

1 . Process for obtaining a plant dichloromethane extract and fraction rich in naphthoquinones from  Eleutherine plicata  plant species characterized by comprising:
 a. Grinding extraction of the sprayed  E. plicata  bulbs(7 days) with ethanol at 96° GL;   b. Concentration in rotatory evaporator under reduced pressure until a residue is formed, being held until a constant weight is obtained;   c. Fractioning of the  E. plicata  bulb ethanol extract (EEBEP) in chromatography column having silica gel as stationary phase and, as mobile phase, the solvents: hexane, dichloromethane, ethyl acetate, and methanol;   d. Concentration of the fraction on a rotatory evaporator under reduced pressure until a residue is formed, being held until a constant weight is obtained.   
     
     
         2 . Process according to  claim 1  characterized by the extract and fractions obtained being additionally purified and presenting an increase on antiplasmodial activity. 
     
     
         3 . Process according to  claim 1  characterized by the extract and fractions obtained having quinones. 
     
     
         4 . Process according to  claim 1  characterized by the dichloromethane fraction having isoleutherine at high purity levels. 
     
     
         5 . Process according to  claim 1  characterized by the ethanol extract and the dichloromethane fraction being active against  P. falciparum  (CI 50 <10 μg/mL), the later being more active; the ethyl acetate fraction moderately active (10 μg/mL<CI 50 <50 μg/mL) and the methanol fraction considerably inactive (CI 50 >50 μg/mL). 
     
     
         6 . Pharmaceutical compositions characterized by comprising the ethanol extract of  Eleutherine plicata  or its dichloromethane fraction and at least a pharmaceutical and pharmacologically acceptable excipient. 
     
     
         7 . Pharmaceutical compositions according to  claim 6  characterized by further comprising an isoleutherine and derivatives thereof. 
     
     
         8 . Pharmaceutical compositions according to  claim 6  characterized by comprising a standardized fraction rich in naphthoquinones and at least a pharmaceutical and pharmacologically acceptable excipient. 
     
     
         9 . Pharmaceutical compositions according to  claim 6  characterized by having antiplasmodial activity. 
     
     
         10 . A method for treatment of malaria comprising administration to an individual in need of treatment for malaria a pharmaceutical composition comprising the ethanol extract of  Eleutherine plicata  or its dichloromethane fraction and at least a pharmaceutical and pharmacologically acceptable excipient. 
     
     
         11 . Method according to  claim 10  characterized by being administered to the individual in need of treatment by oral, intramuscular, intravenous, intraperitoneal, subcutaneous, transdermic routes. 
     
     
         12 . Method according to  claim 10  characterized by alternatively administering the pharmaceutical composition by means of devices able to be implemented or injected.